Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Gonadotropins I and II do not stimulate thein vitro secretion of 17α,20β-dihydroxy-4-pregnen-3-one 20-sulphate by rainbow trout gonads during final sexual maturation

Thein vitro secretion of 17,20-dihydroxy-4-pregnen-3-one 20-sulphate (17,20-P-sulphate) and the free steroid 17,20-dihydroxy-4-pregnen-3-one (17,20-P), by rainbow trout (Oncorhynchus mykiss) gonads, in response to gonadotropin (GTH) I and GTH II, were studied during the final stages of sexual maturation. Substantial amounts of 17,20-P-sulphate were produced, by both mature ovaries and testes, indicating considerable 20-hydroxysteroid sulphotransferase (20-HST) activity within these tissues. In the post-ovulatory ovary the level of 17,20-P-sulphate (36.6 ng ml^–1) greatly exceeded that of 17,20-P (8.59 ng ml^–1). The amount of 17,20-P-sulphate produced in incubations of both mature ovary and testes was unaffected by either GTH I or GTH II treatment at physiological concentrations up to 100 ng ml^–1. Similarly, incubations of maturing ovary and testes, treated with GTH I or GTH II, in the presence of added 17,20-P at 100 ng ml^–1 of medium, produced levels of 17,20-P-sulphate that were similar to those of the controls. In incubations of mature ovarian follicles at the stages of germinal vesicle breakdown and preovulation, both GTHs significantly stimulated secretion of 17,20-P, although GTH II was always more potent than GTH I. GTH II significantly elevated the levels of 17,20-P in testicular incubations from mature males more than 4-fold relative to GTH I and controls, which did not differ from one another.In conclusion, 20-HST, the enzyme responsible for the sulphate conjugation of 17,20-P, was found to be active in the ovaries and testes of rainbow troutin vitro. However, the levels of this enzyme do not appear to be regulated by either GTH I or GTH II.     

Fertility and motility of sperm from Atlantic halibut (Hippoglossus hippoglossus) in relation to dose and timing of gonadotrophin-releasing hormone agonist implant

In broodstocks of Atlantic halibut, Hippoglossus hippoglossus, male and female gamete production often becomes unsynchronised towards the end of the spawning season—milt becomes very viscous and difficult to express while the females are still producing batches of good quality eggs. Gonadotrophin-releasing hormone agonist (GnRHa) has been shown to stimulate spermiation in a number of fish species. Therefore, we conducted two experiments where male halibut were implanted intramuscularly with pellets containing GnRHa. The effect of the pellets was tested at three periods: before, at the height of and at the end of spermiation. In the middle period, GnRHa was tested at two doses (5 and 25 μg/kg bodyweight). Measurements were made of milt hydration, sperm motility and fertilisation rate. Implanted males began spermiation at least 4 weeks before control males. Both doses of GnRHa increased the fluidity of the milt. This effect lasted for at least 20 days in the low dose group and for 40 days in the high dose group. When applied at the end of the season, GnRHa reversed the normal trend for the milt to become more viscous. GnRHa treatments did not affect fertilisation rates obtained with the sperm. However, towards the end of the spawning season, sperm motility was enhanced in males treated with the high dose of GnRHa (25 μg/kg) compared to controls. As described previously, plasma concentrations of the gonadal steroids, 5β-pregnane-3β,17,20β-triol 20-sulphate and 17,20-dihydroxy-4-pregnen-3-one, were significantly enhanced by GnRHa treatment. Concentrations of testosterone on the other hand decreased when spermiating males were treated with GnRHa. Our data suggest that 17,20β-dihydroxy-4-pregnen-3-one or its metabolites are involved in milt hydration, possibly through affecting ion transport.     

HALO AND REVERSE HALO SIGNS IN CANINE PULMONARY COMPUTED TOMOGRAPHY

The halo sign (HS) and reverse halo sign (RHS) are radiologic signs identified on pulmonary computed tomography (CT) in people. The HS is described as a circular area of ground‐glass attenuation surrounding a pulmonary nodule or mass. The RHS is defined as a focal, rounded area of ground‐glass attenuation surrounded by a more or less complete ring of consolidation. These signs have been identified in a variety of diseases in people. The purpose of this retrospective study was to determine if the HS and RHS occur in dogs with pulmonary disease and to determine if they are associated with a particular disease process. In addition, the appearance of the HS and RHS was correlated with the histopathologic changes. Our results indicate that the HS and RHS are not common signs identified in dogs with pulmonary disease with an HS noted in five cases and an RHS in 4 of the 33 dogs that met the inclusion criteria. An association between the HS (P‐value 0.8163) or RHS (P‐value 0.5988) and neoplasia, infectious/inflammatory, and other disease processes was not identified using a Fisher's exact test. The HS was identified in neoplastic, infectious, and inflammatory conditions, with the RHS identified in neoplastic and infectious diseases and a lung lobe torsion. Histologically, the HS and RHS were caused by tumor extension, necrosis, and/or hemorrhage of the pulmonary parenchyma.     

In vitro synthesis of 20α-reduced and of 11- and 21-oxygenated steroids and their sulfates by testes of the goldfish (Carassius auratus): Testicular synthesis of corticosteroids

Testes from spermiating goldfish were incubated with [³H]17-hydroxyprogesterone. The major products in the unconjugated fraction were identified as androstenedione, androstenetrione, 11-β-hydroxyandrostenedione, 11-ketotestosterone, 17,20α-dihydroxy-4-pregnen-3-one (17,20αP) and 11-deoxycortisol. Testosterone was present predominantly in the glucuronide fraction, but yields were low (1–3%). The major components of the sulfate fraction were 17,20αP and 11-deoxycortisol. The identification of cortisone in low yield (< 2.5010) in both the free and sulfate fractions is the first report of corticosteroid biosynthesis by a teleost testis. The high yields of 17,20αP and 11-deoxycortisol and their sulphates suggests that their possible role in spermiation of the goldfish should be further investigated.     

Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.     

Leaf physiology and morphology of Castanea dentata (Marsh.) Borkh., Castanea mollissima Blume,and three backcross breeding generations planted in the southern Appalachians,USA

Backcross breeding programs have been used to transfer disease resistance and other traits from one forest tree species to another in order to meet restoration objectives. Evaluating the field performance of such material is critical for determining the success of breeding programs. In eastern North America, The American Chestnut Foundation has a backcross breeding program that uses Chinese chestnut (Castanea mollissima Blume) to introduce resistance of the fungal pathogen chestnut blight [Cryphonectria parasitica (Murr.) Barr.] to the native American chestnut [Castanea dentata (Marsh.) Borkh.]. We compared physiological and morphological characteristics among seedlings of American chestnut, Chinese chestnut, and BC₁F₃, BC₂F₃, and BC₃F₃ hybrid chestnuts during their fourth growing season after field-planting. American chestnut and the BC₃F₃ breeding generation displayed photosynthetic light-response curves that were similar to each other but different from Chinese chestnut. Rates of photosynthesis were higher for American chestnut and the BC₃F₃ breeding generation when compared to Chinese chestnut for light levels ≥800 μmol m^?2 s^?1 photosynthetic photon flux density and for maximum photosynthetic capacity. Leaf morphology variables were not different between American chestnut and any of the breeding generations, but leaf area (on a per leaf basis) of Chinese chestnut was lower than that of any other chestnut type. Our results suggest that backcross breeding can be used to transfer desirable traits for restoration of native species threatened by non-native pathogens.     

Salinity tolerance during early development of threatened Murray hardyhead (Craterocephalus fluviatilis) to guide environmental watering

1. The worldwide reduction in wetlands has led to the large‐scale decline of wetland‐dependent species. In Australia, to redress some of the decline, partial restoration of the hydrology of a small number of wetlands has been attempted using allocations of environmental water.
2. A common goal of the watering is the maintenance and enhancement of native fish communities, which historically have included populations of the salt tolerant Murray hardyhead (Craterocephalus fluviatilis), a small, short‐lived fish, endemic to the lower Murray–Darling Basin.
3. Despite the addition of environmental water to several sites at which the species is known to persist, populations continue to decline. This decline is, at least in part, suspected to be a consequence of salinities that conflict with the breeding ecology and survival of early life stages.
4. Here the effect of salinity on egg hatch rate and the upper salinity tolerance of larval and juvenile Murray hardyhead was determined under laboratory conditions. It was found that eggs were vulnerable to elevated salinities, whereas juveniles were capable of tolerating salinities up to 105 ppt.
5. Based on the results of the experiment, brackish wetlands managed for Murray hardyhead should be maintained, where possible, between 12 and 45 ppt. Such a salinity regime will necessitate less intensive management of salinity, and a reduced volume of environmental water, providing both environmental and fiscal benefits. The research highlights the benefits of investment in targeted research.